An interdisciplinary approach to reducethe antibiotic load in waterbodies (PharmCycle)

Jörg Andrä2, Falk Beyer3, Gesine Cornelissen2, Jörn Einfeldt1, Jens Heseding1, Kim-Kristin Oelkers1, Klaus Kümmerer4 and Carolin Floeter1

1-3 Hamburg University of Applied Sciences, Faculty Life Sciences, Ulmenliet 20, D-21033 Hamburg, Germany1 Department Environmental Engineering, 2 Department Biotechnology, 3 Department Process Engineering, 4 Leuphana University, Institute of Sustainable and Environmental Chemistry, Scharnhorststr. 1, 21335 Lüneburg, Germany

Abstract

Contact : joerg.andrae_bt@haw-hamburg.de

1. PharmCycle with priority antibioticsExisting antibiotics were selected as priority antibiotics according

to different criteria, e.g. occurrence in the environment, market

volume, mode of action, degradation and toxicity as priority antibiotics.

The priority antibiotics were subject of an enhanced ecotoxicological

risk assessment and wastewater treatment.

2. PharmCycle with sustainable, chemically

processed antibioticsIn the second PharmCycle sustainable, chemical

processed antibiotics will be studied with enhanced

ecotoxicological risk assessment and wastewater treatment

methods.

3. PharmCycle with sustainable,

biotechnologically processed antibioticsIn the third PharmCycle sustainable, biotechnological

processed peptide antibiotics will be investigated with enhanced

ecotoxicological risk assessment and waterwater treatment

methods.

Ciprofloxacin Clarithromycin Doxycyclin Erythromycin Sulfamethoxazol Trimethoprim …

PharmCycles

The interdisciplinary approach of PharmCycle

Sustainable Pharmacy

Sustainable pharmacy considers the

complete life cycle of pharmaceuticals – from

the design, processing, use, disposal to the

occurrence in the environment. It is of high

importance to develop active substances, which

adequately fulfil their purpose and after its use

will be fast and completely degraded in the

environment. This approach focuses on the

primary emission source and considers the

complete life cycle of pharmaceuticals – Benign

by Design. For the chemical design an intensive

research on the transformation products with

experimental and in-silico methods is

undertaken.

Environmental law & management

Recommendations for the implementation of sustainable

pharmacy in environmental law and environmental management

will be proposed.

§

Biotechnological Processing

An antibiotic peptide (Fig. 5) was chosen for biotechnological

production, for which a fast degradation in the environment is

assumed and which already proved antibacterial activity against

e.g. Staphylococcus aureus by maintaining rather low hemolysis.

Corresponding DNA was cloned and expression as a single peptide

or as a peptide-onconase fusion product was achieved in Pichia

pastoris and in E. coli, respectively. Cultivation and peptide

production were performed in a highly instrumented 5 L

bioreactor (Fig. 4). Peptide was purified from inclusion bodies (E.

coli) or from the cultivation supernatant (yeast) by immobilized

metal ion affinity chromatography and/or RP-HPLC (Fig. 6).

Peptide identity was confirmed by mass spectrometry.

Fig. 4: 5 L Bioreactor

Fig. 1: Testorganism

Fig. 6: Purification via RP-HPLC

NK-Lysin

NK-2

(Andrä & Leippe et al. 1999)Fig. 5: Antibiotic Peptide

Fig. 3: Membrane Filtration

Fig. 2: Biological Treatment

Wastewater Treatment

Process steps like biological

wastewater treatment, various

adsorption technologies and

membrane filtration are suitable

individually and in combination

to remove pharmaceuticals from

wastewater.

Among the objectives of

PharmCycle the boundaries of

the removability of

pharmaceuticals from wastewater

and the comparison to the

behavior of sustainable developed

pharmaceuticals in such

treatment steps will be

determined.

The key objective of the research project „PharmCycle“ is to reduce prospective impacts of

pharmaceuticals to the aquatic environment and drinking water supply. PharmCycle addresses therefore

the life cycle of pharmaceuticals, from the sustainable design and production, enhanced ecotoxicological

risk assessment to the improved wastewater treatment.

The first cycle runs with antibiotics, which are selected from different prioritization lists due to their

environmental concern. For these antibiotics an sustainable ecotoxicological risk assessment is performed,

integrating different chronic bacteria tests. A set of wastewater treatment pilot studies aims to reduce the

concentration of the selected antibiotics in effluents.

The second cycle runs with sustainable antibiotics designed with physical-chemical and in silico

methods.

The third cycle is executed with sustainable antibiotics, including newly designed antibiotic peptides,

produced within PharmCycle using molecular and biotechnological methods.

Every cycle includes the upgraded ecotoxicologial risk assessment and investigations on wastewater

treatment procedures. The approach of the project and first results are shown.

Sustainable ecotoxicological risk assessment

The aim is to develop a more sustainable environmental risk assessment of

pharmaceuticals, especially of antibiotics. Therefore further biological test methods

will be developed beside the bioassays required from the EU (EMA) to identify

chronic ecotoxicity, behavioural effects, effects on symbiosis and on marine

(sediment) organisms will be developed. Within PharmCycle a chronic marine

cyanobacteria test was developed, which was significantly more sensitive than the

marine algae test with Phaeodactylum tricornutum according to DIN EN ISO

10253.